RNAs bound by the glucocorticoid receptor during dexamethasone treatment

The RNA binding ability of the glucocorticoid receptor (GR) remains an understudied area of GR regulation. Through in vitro binding assays, we identified hairpin RNAs as GR's preferred binding motif. To study how GR-bound RNAs change with dexamethasone treatment, we first generated stable U2OS cells expressing wild-type GR-HaloTag. We then treated the cells with dexamethsone every hour over three hours, UV crosslinked, harvested, and pulled down GR using a specific and covalently-bound HaloTag ligand. GR-bound RNAs were then isolated, library prepped, and sequenced. RIP-seq with UV crosslinking and HaloTag pulldown to measure changes in the GR-bound RNA population in U2OS cells stably expressing GR-HaloTag over 3 hours of 100 nM dexamethasone treatment.